Table vii



United States Patent Officc- Reissued Mar. 24, 1970 ABSTRACT OF THE DISCLOSURE A method of controlling coccidiosis in animals comprising orally administering to domestic animals an edible carrier containing an effective amount of an anti-coccidial of the formula:

wherein each of R R and R is individually selected from the group consisting of hydrogen, halogen, nitro, lower alkyl, phenyl, and lower alkoxy and R and R or R and R taken together, may form a benzo moiety.

This invention relates to a method of preventing or controlling coccidiosis in domesticated animals such as poultry, turkeys, sheep, goats, cattle, rabbits, etc. More particularly, the present invention resides in the discovery that certain quinazolinone compounds, as described more particularly hereinafter, are highly effective anticoccidials at extremely low dosages.

A broad class of quinazolinones are described in the Baker et al. Patent No. 2,694,711 and are known to be physiologically active as antimalarial agents.

We have discovered that certain of these quinazolinones are effective anticoccidials at very low concentrations. This is surprising because it is known that little or no correlation exists between useful antimalarial activity and useful anticoccidial activity. So far as we are aware no useful anticoccidial is a useful antimalarial and no useful antimalarial is a useful anticoccidial; therefore, it is surprising to discover that a limited number of the Baker et al. quinazolinones are exceptionally active at conspicuouly low concentrations, i.e., 0.5 to 10 p.p.m. Such exceptional activity is required for practicality as an anticoccidial as the synthesis of this class of compounds is complex and expensive. Furthermore, We have found a limited number of the Baker et al. quinazolinones to be highly ettective against a wide variety of coccidial species including E. tenella, E. necatrix, E. acel'vulina, E. meleagrimitis, E. maxima and E. brunetti.

The active anticoccidial agents of the present invention are compounds represented by the general formula:

Rt R) l t 1 N-ciIt-d-c1n wherein each of R R and R is individually selected from the group consisting of hydrogen, halogen, nitro, lower alkyl, phenyl and lower alkoxy, and R and R or R and R taken together, may form a benzo moiety. [wherein R is a member of the group consisting of hydrogen, halogen, nitro, benzo, lower alkyl, phenyl and lower alkoxy,] R is a member of the group consisting of hydroxy, acetoxy, and lower alkoxy and R is a member of the group consisting of hydrogen and lower alkenoxycarbonyl. Suitable lower alkyl and lower alkoxy radicals are those having from 1-6 carbon atoms. [The substituents represented by the symbol R may be one or more of the 6, 7, or 8 positions only of the quinazolinone nucleus] Also included with the scope of the present invention are the salts of these compounds with either one or two equivalents of a physiologically acceptable acid.

Although for the past decades coccidiosis has been recognized as one of the most important diseases confronting the poultry industry, nevertheless, heretofore, no entirely satisfactory method of control of the disease has been provided. This is evidenced by the fact that losses in the United States due to poultry coccidia are estimated to exceed 38 million dollars annually. Coccidiosis is also an important disease of sheep, goats, cattle, rabbits, etc.

Many know anticoccidials have shown high activity against E. tenella and E. necatrix, but low activity against other important species of coccidia, such as E. acervulina, E. meleagrimitis, E. maxima and E. brunetti. Furthermore, the commercially available anticoccidials require relatively high concentrations in the animal ration, i.e., 50 and frequently more parts per million, whereas the compounds of the present invention will effectively control coccidiosis at 0.5 to 10 p.p.m.

The present invention involves the method of preventing, controlling or treating coccidiosis involving orally administering to animals either prophylactically or therapeutically an effective amount, i.e., in the range of 0.5 to p.p.m. of one of the compounds of the above formula. Generally it has been found that between about 0.5 and 10 p.p.m. of one of the active anticoccidials of the present invention in the diet of the animal or in the drinking water is highly effective in preventing or controlling or treating coccidiosis.

When using the compounds of the invention for the prevention or control of coccidiosis, the active coccidiostat is normally first compounded into an animal feed supplement. These supplements, which contain relatively large percentage of the coccidiostat, are then uniformly distributed in the finished animal feed either directly or after an intermediate processing step. The feed supplements are prepared by adding the subject compounds to a suitable carrier and mixing for sufiicient time to give substantially uniform dispersion of the coccidiostat in the carrier. These supplements ordinarily contain about (ll-1.0% and preferably from about 0.30.6% of active ingredient. The carrier or diluent is a solid edible material that is inert with respect to the subject compounds and that may be administered with safety to the animals to be treated. Typical of such carriers are distillers grains, corn meal, ground oyster shells, attapulgus clay, molasses solubles, antibiotic mycelia, soya grits, soyabean feed, soyabean meal, crushed limestone and the like. For the preparation of these supplements, fats, oils, antioxidants, and surface active agents can be employed.

These highly active anticoccidials may be combined with other anticoccidials such as the sulfonamides, or with feed additives, such as antibiotics, vitamins, etc.

The invention will be described in greater detail in conjunction with the following specific examples.

Example 1.E. tenella-Prophylactie In the following tests, 1125 Rhode Island Red male x Barred Rock female cross cockerels, 7 days old and of approximately equal size and weight, were divided into eleven groups comprising from 10 to 279 chicks per group. These groups were placed in separate cages with wire floors. A broiler feed diet of the formulation set forth below and having graded levels of test medicament intimately blended therewith was prepared and administered to the various groups. The formulation used was as follows:

FORMULATION Lbs/ton Corn yellow, fine ground 1200 Soybean, fine ground 400 Corn gluten meal 100 Fish meal 100 Alfalfa meal 40 Distillers solubles 50 Mico mix 8009 Limecrest-t-MnSO 40 Bone meal (steamed) 30 NaCl 10 Choline chloride 2 Fortafeed (249C) 2 4 g. pantothenie/lb.

9 g. niacin.

10 g. choline. Vitamin A+D, dry 2 10,000 ,u/g. A.

Profactor B, 10 mg./ton.

The medicaments employed in the instant tests were 4(3H)-quinazolinone, 6,8dichloro 3-[3-(3-hydroxy-2- piperidyl)-acetonyll-, hydrochloride; 4(3H)-quinazolinone, 6 phenyl 3-[3-(3-hydroxy-2-piperidyl)acetonyl]-, dihydrochloride and 4(3H)-quinazolinone. 7-bromo-6'chloro- 3-[3-(3-hydroxy-2-piperidyl) acetonyl], hydrobromide.

The medicated and unmedicated diets were presented to the chicks and the chicks were permitted to feed and drink ad libitum from two days prior until 7 days following the oral inoculation with sporulated oocysts of Eimeria tenella. The number of oocysts inoeculated directly into the crops of all chicks in the test was sufficient to produce 8S100% mortality in the untreated controls. The quantity necessary to produce this mortality rate was determined prior to the time of inoculation by giving graded quantities of oocysts to comparable birds. Seven days following inoculation the test was terminated and the mortality rate recorded for each group.

The results obtained are provided in Table I below where it can be seen that as little as 0.5 part per million of 4(3H)-quinazolinone, 6,8-dichloro-3-[3-(3-hydroxy-2- piperidyl)acetonyH-hydrochloride reduced the mortality rate in chicks infected with the organism E. tenella and 5 p.p.m. afforded complete protection against the organism. It may also be noted that as little as 2 p.p.m. of 4(3H)-quinazolinone, 7-bromo-6-chloro-3-[S-(Z-hydroxy- 2-piperdyl)acetonyl]-hydrobromide and 2.5 p.p.m. of 4(3H)-quinazolinone, 6-phenyl-3-[3-(3-hydroxy-2-piper idyl)acetonyl]-dihydrochloride reduced the mortality rate of chicks infested with E. tenella and 4 and 10 p.p.m. respectively produced 100% survival. Although the formulation given above was used in the above tests, other standard animal feed formulations can be used with similar success.

TABLE I.ANTICCOCIDTALPROPIIYLACTICE.

TENELLA Example 2.E. tenellaTherapeutic Seven-day-old Rhode Island Red male x Barred Rock female cross cockerels of substantially similar size and weight were separated into groups of 20, 40, and 50 birds. The birds in each group were orally inoculated with a sufficient number of sporulated oocysts of E. tenella to produce l00% mortality in untreated controls. As in Example I, the oocyst quantity was determined prior to test inoculation. Following inoculation the groups were separately caged in units with wire floors beneath which were placed white papers to catch the droppings and make daily visual examination thereof convenient. For the first three days following inoculation, all groups were fed ad libitum, a broiler feed diet of the formulation given in Example 1 with no medication added. At 72 hours after inoculation, shortly before blood appeared in the droppings, 1.5, 2.0, and 0.0 p.p.m. of 4(3H)-quinazolinone, 6,8-dichloro-3-[3-(3-hydroxy-2-piperidyl)acctonyl] hydrochloride were thoroughly mixed with the feed formulation and presented to the groups of 20, 40, and 50 birds, respectively. Medication was continued for four days and then the test was terminated and the number of survivors in each group counted and recorded.

Further, employing the above procedure but administering the drug in 2000 ml. of drinking water instead of in the feed produced results similar to those obtained with medicated feed.

The results appear in Table II below where it can be seen that only 2.0% of the birds receiving unmedicated diet survived, whereas 80% of those receiving the diet containing 2 ppm. of the medicament survived. With water medication where only 25% of the unmedicated birds survived, 100% of the birds medicated with 2 p.p.m. survived. The therapeutic effect of the medicament against E. tenella is, therefore, apparent.

TABLE II.E. TENELLATHERAPEUTIC ACTIVITY vidually weighed and returned to their cages. A broiler feed diet of the formulation set forth in Example 1, but having graded levels of medicament intimately dispersed therein, was administered to the several groups of birds for the entire duration of the test. Two days following presentation of the test diets, all birds were orally inoculated with a sufficient number of sporulated ooeysts of E. necatrix to produce severe intestinal lesions and 80% mortality in the untreated control group. As in previous examples, the number of ooeysts required to produce the desired extent of infection was determined prior to the time of inoculation with comparable birds. Seven days after inoculation the test was terminated. The number of survivors in each test group was recorded, each bird Parts per Number Percent was individually weighed and the average percent weight Compound million 52 3 survival gain for each group calculated and recorded. Following weighing, all birds were necropsied and the number in Feed medication: each group with reduced intestinal lesions determined by liiigl-qfinazo linpnafiS-fiehlordlig 40 an examination of the intestinal tracts. Criteria for activity l g gg g p p y t y l-, g g were percent survival, the number of birds with markedly reduced lesions in the small intestine (fewer than 100 Inoculated pinpont lesions localized immediately around the yolk wamrmedicafiom stalk) and weight gains made relative to the uninfected 5 %;q5 9 .f ichlow?- 3 1% unmedicated controls. Lesions 1n the untreated controls L gg ggfiggf y me my 0 20 25 were so abundant and extensive as to be uncountable and produced a very severe hemorrhagic enteritis. Uninoeulated From Table III below it is evident that as little as 1.0

ppm. of these compounds, when administered prophy- 0 20 100 lactically, will effectively control E. necatrix, the causative agent for intestinal coccidiosis in chicks.

TABLE III.NECATRIXPROPHYLACTIC Number Parts per Number with Avg. per- Compound million birds Percent reduced eent wt.

in diet treated survival lesions gains Uninieeted 0 20 100 112 Infected (1) 20 20 0 20 0 39 4(3H)-qulnazollnone, 6, 8-diehloro-3-[3-(hydroxy-Z-piperidyl)aeetonyl}. hydrochloride. g 33 33 g 4(3H)-quinazolln0ne, 6, 7-diehloro-3-[3-(3 1 20 S5 1 69 hydroxy-2-piperidyl)aeetonyl]-, hydrobro- 1. 5 20 3 104 rnide. 25 3 100 19 11s 0 0 e1 4(3H)-qulnazolinone, 6-ehloro-3-[3(3-hydroxyg 2-piperldyl)acetonyll-, hydrobromide. 3 g Unlnieeted 0 40 100 131 Infected 0 40 20 0 4(3H)-quinazolir1one, 7,8dichlert 3-[3-(3- 4 20 9 122 hydroxy-Z-piperidyl)aeetonyl]-, hydrobro' 5 40 97 31 118 mlde. 6 20 18 119 Example 3.E. necatrix-Prophylactic Example 4.E. maximaProphylactic To determine the effectiveness of the compounds against the infective organism E. maxima, which is responsive for one type of intestinal coccidiosis in chickens, Rhode Island Red male x Barred Rock female cross cockerels were randomly divided into three equal groups. The chicks in all groups were individually weighed and the groups placed in separate cages with Wire floors. A broiler feed diet of the formulation described in Example 1 and having intimately dispersed therein graded levels of 4(3H)-quinazolinone, 6,8-dichloro-3[3-(3-hydroxy 2-piperidyl)acetonyl]-, hydrochloride and 4(3H)- quinazolinone, 6,7 dichloro 3[3-(3-hydroxy-2-piperidyl)acetonyl]-, hydrobiomicle were provided for the various groups. The chicks were permitted to feed ad libitum throughout the entire test. Two days after presentation TABLE AGERVULINA A PHOPHYLACTIO of the test feeds all chicks were inoculated with approxi- Parts per Number Avg.percent mately 150,000 sporulated oocysts of the organism E. Compound minionin birds Weigmgam maxima. Ten days after inoculation the chicks were ret treated uninfected I" moved from their ca es Wei hed the we1 hts recorded g g Uninfeeted 0 20 100 and the test was terminated. in r fiem nn 1 53 gg 4 unazoinone, In a similar test with 100 chicks d1v1ded into groups 3. m 20 s id -h of 20 birds each, all chicks were inoculated with 40,000 10 gl fg gi 8 3g 4 3H i an ll 0 e,6,7-disporulated oocysts after 2 days of feeding graded levels zgf fl 1 20 75 of 4(3H) quinazolinone, 7-bromo-6-chloro-3-[3-(3-hygigfggg 20 93 4(3H)quinazolinone,6'chloro 1 20 91 droxy 2 piperidyhacetonyll, hydrobromide. 3%(mhydmxyvzmipmdyly 1. 5 20 94 The results are tabulated in Table IV below from which q y l-, v d i Umnlected 0 40 100 it is evident that the test compounds were effective in con- Infected..." 0 40 72 4(311) ouinazollnoneJ-bromo U'Olllng E. maxima at a concentration in the feed of as eehioreaatah dmx aa 30 97 i d 1- l-,h d 4 102 little as 1 or 2 ppm. 8,231. 5) acuwny] y m- TABLE IV.-E. MAXIMAPROPHYLACTIC Grams Parts per Number gained million birds Percent per bird Compound in diet treated survival 10 days Uniniccted 0 20 100 124 Infected 0 20 05 90 4(3ll)-qninazollnone, ti, 8-dichloro-3l3- 1 20 100 103 (341ydroxy-Q-pipcridyl)acetonyl]-, 1. 5 20 100 110 hydrochloride. 2. 20 100 108 4t3H)-quinuzolinone, 6,7-dlchloro-3-{3- 1 20 100 105 til-hydroxy-Z-piperidyl)acetonyl]-, 1. 5 20 100 109 hydrobrornldc. Uninfected 0 20 100 184 Infected. 0 20 100 128 4(3ll)-quinazolinone, 7-brom06-chlor0- 2 20 100 159 3-[3-(S-hydroxy-Z-plperidyh- 3 20 100 187 acetonyl} hydrobrornide. 4 20 100 186 Example 5.E. acervulina-Prophylactic In determining the efiicacy of the compounds against the infective organism E. acervulina (one of the etiological agents causing intestinal coccidiosis in chickens), 200 Rhode Island Red male x Barred Rock female cross cockerels, 7 days old, were randomly divided into 10 groups each comprising 20 chicks. These groups were caged in separate units and given free access to feed and water. The diet employed was a broiler feed of the formulation given in Example 1 to which had been added graded levels (from 0 to 3.37 ppm.) of 4(3H)-quinazolinone, 6,8-dichl0ro 3-[3-(3-hydroxy 2 piperidyl)acetonyl]-, hydrochloride, and 0 to 1.5 ppm. of 4(3H)- quinazolinone, 6,7-dichloro 3-[3-(3-hydroxy Z-piperidyl)acetonyl]-, hydrobromide and 4(3H)-quinazolinone, 6-chloro-3-[3-(3-hydroxy-2-piperidyl)-acetonyl]-, hydrobromide. Two days after presentation of the prescribed diets to the various groups, all chicks were orally inocculated with about 600,000 sporulated oocysts of E. acervulina. The inoculated birds were returned to their cages and the diet was continued for 10 days until the experiment was terminated. On termination, all birds were removed from their cages and weighed, and the weights were recorded.

In a similar test with 130 chicks divided into groups of 2-0 to chicks each, all chicks were inoculated with 700,000 sporulated oocysts after 2 days of feeding graded levels of 4(3H)-quinazolinone, 7-bromo-6-chloro-3-[3- (3-hydroxy-2-piperidyl)acetonyH- hydrobrornide.

The results of the test indicate that as little as 1 ppm. of the quinazolinones tested furnished substantial protection for the birds against the organism E. acervulina. The results of the test recorded in Table V above.

Example 6.-E. brunettiProphylactic In determining the efficacy of the quinazolinones invention against the infective organism E. brunetti, Rhode Island Red male x Barred Rock female cross cockerels, 7 days old, were randomly divided into 5 groups comprising 20 chicks. These groups were caged in separate units and given free access to feed and water. The diet employed was a broiler feed of the formulation given in Example 1 to which had been added graded levels of 4( 3H -quinazolinone, 6,7-dichloro-3- 3- 3-hydroxy2-piperidyl)aeetonyl]- hydrobromide. Two days after presentation of the prescribed diet to the various groups, all chicks were orally inoculated with about 140,000 sporulated oocysts of E. brunetti. The inoculated birds were returned to their cages and the diet was continued for 10 days until the experiment was terminated. On termination, all birds were removed from their cages and weighed, and the weights were recorded.

The results of the test indicate that as little as 1 ppm. of the quinazolinone tested furnished substantial protection for the birds against the organism E. brunetti. The results of the test are recorded in Table VI below.

Example 7 Employing the procedure, formulation and using the test organism, E. tenella, as in Example 1, graded levels of the following compounds Example 8.E. meleagrimitisProphylactic In determining the efficacy of the quinazolinone compounds against the infective organism E. melcagrimitis 29 broad breasted white male turkey poults, 3 weeks old,

5 were randomly divided into 3 groups comprising 11, 9, S and 9 oults, respectively. These groups were caged in N\ p d r r a d 7 separate units an given ree access to ee an water. (111} i The diet employed was a broiler feed of the formulation 6 NTCHPCOTCHP N salt 10 given in Example 1 to which had been added graded l levels of 4(3H)-quinazolinone, 6,8-dichloro-3-[3-(3-hy- 5 R droxy-Z-prpendyl)-acetonyl]-, hydrochloride and 4(3H)- quinazolinone, 6,7-dichloro-3-[3-(3-hydroxy-2-piperidyl) R! acetonyl]-, hydrobromide. One day after presentation R of the prescribed diets to the various groups, all poults 0 were orally inoculated with about 500,000 sporulated J I g -Salt oocysts of E. meleagrimitis. The inoculated birds were R returned to their cages and the diet was continued for H k 9 days until the experiment was terminated. On termina O tion, all birds were removed from their cages and weighed, were administered in the diet and the minimum level of gg weilgthts i q gi t th t rm 1 5 compound necessary to atford protection against mortality 6 g O f a l as of chickens due to the organism determined. The ret e qumazp none .[este su smnha sults obtained in these tests are provided in Table VII proleqtlon for the buds agamst the orgamsm male" balow agrimitis. The results of the test are recorded In Table III below TABLE v11 V Anticoccidial activity against the ineffective organism E. tenella of compounds of the general formula:

s r N\ 2 TABLE VIII.E. MELEAGRIMITIS-PROPHYLACTIC 6 NCH2COCHz .Salt

N Partsper Number Percent Grams 1L Compound million birds survival gained a in diet treated per bird 10 days N Inicc ted 0 1t 91 396 R R 40 4(3H)-quinazolinone,6,8-

W I dichloro-Jl-[3-(3-hydr0xy- H S u Zptpendyllacetonyllw R N-cHr-c'-cH,-' hydrochloride 2 9 100 453 N 4(3H)-quinazolinone, 6,7- I I dichloroQ-[B-(ll-hydroxy- R 2-piperidyl)acetonyl]-,

hydrobromide 1. 5 9 100 481 Parts per R5 R7 R5 R2 R3 Salt million in diet 1 H U 1 H 3 2 g g 2 Example 9.--Mrxed coccidia infection of E. tenella and H m E. acervulina H HCl 10 g 3 A mixed inoculum of 5000 sporulated oocysts of E. g l l i3 acervulina and a sufficient number of oocysts of E. tenel- H 1 la to produce 85-100% mortality in untreated controls gg g was given to seven-day-old chicks, similar to those de- H 2 scribed in previous tests. The chicks were given free acg cess to feed and water during the entire test period. Two days after inoculation medicated feed, with several H 2110! 10 g levels of drug or drugs, was presented to the various H 1 groups in test. Ten da s after inoculation the tests were H gm?! 2 d d b b' d 'hd 'd d terminate an t e 1r s were welg e necropsie an H 10 their intestinal tracts examined for lesions. The results H 1131 a g i of the tests appear in Table IX. These data show that H HBr 4 mixtures of quinazolinones and a sulfonarnide such as 6 CH H01 i3 sulfaethoxypyridazine (i.e., 3-sulfanilamido-6-ethoxypycli o co ridazlne) give complete protection against a mixed coc- 2H9! i3 ctdla infection at relatively low levels. The data also show that the sulfonamide alone alfords no protection H BB 2 against E. tenella lesions at any level used and that the r addition of a sulfonamide to a feed medicated with a quin [R wlth equalsH azolinone reduces the amount of said material necessary RBJRT and Rs with" equal H, to achieve desired control.

TABLE IX I Sulfnethoxy- Number of Number Percent Supression Compound, ppm. in diet pyridazine birds in test survivors lesion E. acerulina.

group E. tenella 6,8-dlchloro-3-[3-(3-hydroxy-2 piperidyl)-acetonyl]4,(3H)- quinuzolinone:

2 60 10 100 100 30 50 50 84 94 50 47 80 86 0 60 59 83 47 60 20 70 100 0 20 19 65 45 0 20 20 80 100 250 10 9 0 100 0!) 10 3 0 100 t) O 80 12 0 0 7,8 dichlorrr3-[3-(3-hydroxy-QF piperidyl)acetonyl]-,4(3ll)- quinazolinone:

Example 10.Preparation of allyl-2-(3-bromoacetony1)- 3-methoxyl-piperidine carboxylate This compound is prepared by the method of B. R. Baker and F. J. McEvoy, J. Org. Chem. 20, 141 (1955), using 2-acetonyl-3-methoxypiperidine synthesized as described in the Barringer and Berkelhammer copending application filed concurrently herewith.

Example 11.Preparation of 6,7,8-trichloro-4(3H)- quinazolinone 3,4,5-trichloroanthranilic acid (8.6 grams) and 6.3 grams of formamide are dissolved in 20 ml. of dimethylformamide. The solution is heated at reflux for 16 hours. The product crystallizes upon cooling, is collected by filtration, and washed with ethanol. Recrystallization from methyl Cellosolve gives 2.5 grams (30%) of yellow crystals, melting point 298-300".

Example 12.-Preparation of 6,7,8-trichloro-3-[3-(l-allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl]- 4(3H)-quinazolinone Example l3.Preparation of 6,7,8-trichloro-3-[3 (3-hydroxy 2 piperidyl)acetonyl] 4(3H) quinazolinone hydrobromide The product from Example 12 is dissolved in 75 ml. of 48% hydrobromic acid and heated at reflux for 1 hour. The solution is evaporated under vacuum to dryness. The residue is heated for 30 minutes in refluxing 2,8-ethanol (remains undissolved), then collected by filtration; yield 0.85 gram (29%) of crude product. Recrystallization from ethanol-water (activated charcoal) gives white crystals, melting point 239 dec.

Example I4.Preparation of 7,8-dichloro-4( 3H quinazolinone The procedure of Example 11 is followed, using 41.85 grams of 3,4-dichloroanthranilic acid [P. W. Sadler and R. L. Warren, J. Am. Chem. Soc., 78, 1251 (1956)] and 36 grams of formamide in 100 ml. of dimethylformamide. The product is purified by recrystallization from dimethylformamide and washing with acetone. 20.4 grams (50%) of pale yellow crystals are obtained, melting point 287.

Example 15.Preparation of 7,8-dichloro-3-[3-(l-allyloxycarbonyl 3 methoxy 2 pipen'dyl)acetonyl]- 4 3 H) -quinazolinone The procedure of the twelfth example is followed, using 1.29 grams of 7,8-dichloro-4(3H)-quinazolinone and 2.0 grams of a1lyl-2-(3-bromoacetonyl)-3-methoxy-l-piperidine carboxylate. 2.4 grams (86%) of product is obtained as a viscous, amber oil.

Example 16.Preparation of 7,8-dichloro-3-[3-(3-hydroxy 2 piperidyl)acetonyl] 4(3H) quinazolinone hydrobromide The product from Example 15 is hydrolyzed following the procedure of Example 13. In this reaction a homogeneous solution is formed when refluxing the residue with ethanol. The solution is evaporated under vacuum and the residue triturated with cold ethanol. The brown solid is collected by filtration, then recrystallized from ethanolwater (activated charcoal). 340 mg. of product is obtained as a While crystalline material, melting point 248 dec.

Example 17 The compounds listed below, previously known as the dihydrochloride salts, are herein prepared according to Examples 12 and 13:

6-chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl]-4(3H)- quinazolinone hydrobromide (melting point 240 dec.)

6-bromo-3- 3-(3-hydroxy-2-piperidyl acetonyl] -4( 3H quinazolinone hydrobromide (melting point 257 dec.)

Example 18 The following compounds, previously known as the dihydrochloride salts, are prepared as the doubly blocked condensation products as in Example 12. The allyloxycarbonyl blocking group is removed by heating the crude condensation product in 50 ml. of 6 N hydrochloric acid at steam-bath temperature for 1 hour and evaporating to dryness. The methyl ether is then hydrolyzed by treating this residue as in Example 13.

8-chloro-3-[3- (3-hydroxy-2-piperidyl)acetonyl] -4(3H) quinazolinone hydrobromide (melting point 247 dec.) 6,7-dichloro-3- 3- 3 -hydroxy-2-piperidyl acetonyl] 4( 3H) -quinazolinone hydrobromide (melting point 237 dec.) 6,8-dichloro-3-[3-(3-hydroxy-2-piperidyl acetonyl] 4( 3H)-quinazolinone hydrobromide (melting point 25 1 dec.

1 3 Example 19 The free base of 6-chloro-3-[3-(3-hydroxy-2-piperidyl)- acetonyl]-4(3H)-quinazolinone, previously known as the dihydrochloride, is prepared by neutralizing the hydrobromide (prepared as in Example 17) with 1 N sodium hydroxide and extracting into chloroform. Evaporation of the chloroform leaves the free base as a white solid, melting point 176-177".

Example 20.Preparation of 6,7-dichloro-3-[3-(l-allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl]- 4(3H)-quinazolinone The procedure of Example 12 is followed, using 1.29 grams of 6,7-dichloro-4(3H)-quinazoline [B. R. Baker et al., J. Org. Chem, 17, 149-56 (1952)] and 2.0 grams of allyl-2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. 2.3 grams (82%) of a viscous oil is obtained which is chromatographically purified to yield a white crystalline product, melting point 101-102.

Example 21.Preparation of 6,7-dichloro-3-[3-(3-acetoxy-2-pipe ridyl acetonyl] -4( 3H -quinazolinone Thionyl chloride (1 m1.) is added to 400 mg. of 6,7-dichloro 3[3 (3 hydroxy 2 piperidyl)acetonyl]- 4(3H)-quinazolinone hydrobromide suspended in a mixture of 1 ml. of acetic anhydride and 1 ml. of glacial acetic acid. A homogeneous solution is formed accompanied by vigorous evolution of sulfur dioxide. The white crystalline product precipitates upon standing, is removed by filtration, and is washed with ether. The etherwashed solid is dissolved in chloroform and the solution washed thoroughly with aqueous sodium carbonate. The chloroform layer is separated, dried, and evaporated under vacuum, yielding an oil which separates as a white crystalline solid (60 mg.) M.P. 150151 after solution in refluxing hexane-acetone and cooling.

Example 22.Preparation of 6,8-diiod 3-[3-(l-allyloxycarbonyl 3 methoxy 2 piperidyDacetonyl] 4(3H) quinazolinone The procedure of Example 12 is followed, using 2.39 grams of 6.8diiodo-4(3H) quinazolinone [M. R. Subbaram, J. Madras Uni., 248, 17982 (1954); C. A. 50, 3522 (1956)] and 2.0 grams allyl-2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. In this reaction part of the product precipitates from the reaction mixture, is collected by filtration and recrystallized from ethanol yielding 1.23 grams of white crystals, melting point l54l56. The remainder of the reaction mixture is worked up in the usual manner, yielding the usual viscous, oily product.

Example 23.-Preparation of 6,8-diiodo-3-[3-(3-hydroxy- 2 piperidyl)acetonyl] 4(3H) quinazolinone hydrobromide The product of Example 22 is hydrolyzed according to the procedure of Example 13. 250 mg. of White crystalline product is obtained, melting point 244 C. dec.

Example 24.-Preparation of 6-nitro-3-[3-(l-allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl] 4(3H)- quinazolinone The procedure of Example 12 is followed, using 1.15 grams of 6-nitro-4(3H)-quinazolin0ne and 2.0 grams of allyl-2(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate, except that no excess sodium methoxide is added. The product is a viscous brown syrup.

Example 25.Preparation of 6-nitro-3-[3-(3-hydroxy-2- piperidyl)acetonyl]-4(3H)-quinazolinone hydrochloride The product from Example 24 is hydrolyzed according to the procedure of Example 13. The residue remaining from the removal of the hydrobromic acid is treated with refluxing saturated ethanolic hydrochloric acid for 30 minutes and the resultant solid filtered 011 and dissolved in aqueous ethanol. The ethanolic solution is heated under reflux for 30 minutes, treated with decolorizing carbon, and filtered. Refrigeration of the filtrate gives 610 mg. of the hydrochloride salt as pale yellow crystals, melting point 215 dec.

Example 26.Preparation of 6,S-dibromo-4(3H)-quinazolinone The procedure of Example 11 is followed using 0.9 gram of formamide and 2.95 grams of 3,5-dibromoanthranilic acid in 10 cc. of dimethylformamide. The product is purified by slurrying in hot ethanol and filtered to give 2.45 grams (82%) of pale yellow needles, unmelted at 300.

Example 27.Preparation of 6,8-dibromo-3-[3-(l-allyloxycarbonyl 3-methoxy-2-piperidyl)acetonyl]-4(3H)- quinazolinone The procedure of Example 12 is followed, using 1.82 grams of 6,8 dibromo 4(3H)-quinazolinone and 2.0 grams of allyl-2-(3-bromoacetonyl)-3-methoxy-l-piperidine carboxylate, except that no excess sodium methoxide is used. The product is a dark brown syrup.

Example 28.Preparation of 6,8-dibromo3-[3-(3-hydroxy 2-piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 27 is heated under reflux with 30 cc. of 6 N hydrochloric acid for 1 hour and the solution evaporated to dryness. The residue is dissolved in 20 cc. of 48% hydrobromic acid and the solution heated under reflux for 30 minutes and evaporated to dryness. The brown residue is treated with boiling saturated ethanolic hydrochloric acid for 20 minutes, and the resulting solid collected by filtration. In order to insure complete removal of the methyl ether blocking group, the material is subject to rehydrolysis following the procedure of Example 13. 360 mg. of the crystalline hydrobromide is obtained, melting point 250 dec.

Example 29.-Preparation of 6-fluoro-4(3H)-quinazolinone The procedure of Example 11 is followed, using 2 cc. of formamide and 1.1 grams of S-fluoroanthranilic acid in 3 cc. of dimethylformamide. Recrystallization of the crude mass from ethanol gives 550 mg. of colorless product, melting point 249-251 Example 30.Preparation of 6-fluoro-3-[3-(l-allyloxycarbonyl 3 methoxy 2-piperidyl)acetonyl]-4(3H)- quinazolinone The procedure of Example 12 is followed, using 0.55 gram of 6 fluoro-4(3H)-quinazolinone and 1.47 grams of 2-(3-bromoacetonyl)-3-methoxy-1-piperidyl carboxylate, except that no excess sodium methoxide is added. The product is a brown viscous syrup.

Example 3l.-Preparation of 6-fiuoro-3-[3-(3-hydroxy- Z-piperidyl)acetonyl]-4(3H) quinazolinone hydrobromide The product of Example 30 is hydrolyzed following the procedure of Example 13. The brown solid is recrystallized from 99% ethanol (decoiorizing carbon) to give 253 mg. of product, melting point 230 dec.

Example 32 The citrate salt of 6 chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl]-4(3H)-quinazolinone is obtained by taking up the 336 mg. of the free base of this compound (see Example 19) in ethanol and adding an ethanolic solution of 192 mg. of citric acid. Evaporation of the solvent gives the white crystalline citrate.

15 Example Iii-Preparation of S bromo-6-chloro-3-[3-(1- allyoxycarbonyl 3 methoxy-Z-piperidyl)acetonly]-4 (3H)-quinazolinone The procedure of Example 12 is followed, using 3.1 grams of 8bromo-6-chloro-4(3H)-quinazolinone and 4.0 grams of allyl 2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. The product is a viscous amber oil.

Example 34.Preparation of 8 bromo-6-chloro-3-[3-(3- hydroxy 2 piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 33 is hydrolyzed following the procedure of Example 13. A white crystalline product is obtained, melting point 250 dec.

Example 35.Preparation of 6-bromo-8-chloro-3-[3-(1- allyloxycarbonyl 3 methoxy 2-piperidyl)acetonyl]- 4(3H)-quinazolinone The procedure of Example 12 is followed, using 1.55 grams of 6-bromo-8-chloro-4(3H)-quinazolinone and 2.0 grams of allyl 2-(3-brornoacetonyl)-3methoxy-l-piperidine carboxylate. 2.5 grams (85%) of product is obtained as a viscous amber oil.

Example 36.-Preparation of 6-bromo-8-chloro-3-[3- (3 hydroxy 2 piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 35 is hydrolyzed following the procedure of Example 13. 0.8 gram of crude product is obtained which recrystallizes from ethanol-water (activated charcoal) as a white crystalline material, melting point 251 dec.

Example 37.Preparation of 6-bromo-7-chl0ro-4(3H)- quinazolinone 4-chloroanthranilic acid (17.2 grams) is dissolved in 100 ml. of p-dioxane. After heating the solution to 60, 16 grams of bromine dissolved in 100 ml. of p-dioxane is slowly added with stirring. The product precipitates as a yellow solid. It is dissolved in base and reprecipitated with acid, then recrystallized from ethanol.

5.4 grams of this anthranilic acid is cyclized as in Example 11, using 25 ml. of formamide and 10 ml. of dimethylformamide. The product is purified by recrystallization from ethyl cellosolve and washing with ethanol. Light tan needles are obtained, melting point 290-95.

Example 38.Preparation of 6-bromo-7-chloro-3-[3-(1- allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl]- 4(3H)quinazolinone The procedure of Example 12 is followed, using 1.55 grams of 6-brorno-7-chloro-4(3H)-quinazolinone and 2.0 grams of allyl 2-(3-bromoacetonyl)-3-methoxy-l-piperidine carboxylate. The product is a viscous amber oil.

Example 39.Preparation of 6-bromo-7-chloro-3-[3-(3- hydroxy 2 piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 38 is hydrolyzed following the procedure of Example 13. A white crystalline product is obtained, melting point 242 dec.

Example 40.Preparation of 7-bromo-6-chloro-4(3H)- quinazolinone 6-bromoisatin (12.0 grams) is dissolved in 300 ml. of glacial acetic acid, heated to 80, and 10 ml. of sulfuryl chloride added. After 30 minutes the solution is cooled to 20, causing 7.2 grams (55%) of 6-bromo-5-chloroisatin to separate as deep red crystals, melting point 284- 285". The isatin is oxidized with 30% hydrogen peroxide in N NaOH giving 5.8 grams (84%) of -bromo-4-chloroanthranilic acid as tan crystals, melting point 251-252.

5.0 grams of this anthranilic acid is cyclized as in Example 11 using ml. of formamide and 10 ml. of dimethylformamide. The addition of 150 ml. of ethanol to the reaction mixture causes the separation of 3.2 grams (60%) of tan crystalline product, melting point 290.

Example 41.-Preparation of 7-bromo-6-chloro-3-[3-(1- allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl]- 4 3H -quinazolinone The procedure of Example 12 is followed using 2.95 grams of 7-bromo-6-chloro-4(3H)-quinazolinone and 3.34 grams of allyl 2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. The product is a brown syrup.

Example 42.Preparation of 7-bromo-6-chloro-3-[3-(3- hydroxy 2 piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 41 is hydrolyzed following the procedure of Example 13. 410 mg. of tan crystalline product is obtained, melting point 247 dec.

Example 43. Preparation of 6,7-dibromo-4(3H)- quinazolinone 4.0 grams of 4,5-dibromoanthranilic acid is cyclized as in Example 11 using 4 ml. of formamide and 8 ml. of dimethyl formamide. Recrystallization from ethanol-dimethylformamide (10:1) gives 3.9 grams of tan crystals, melting point 310.

Example 44.Preparation of 6,7-dibromo-3-[3-(l-allyloxycarbonyl 3 -methoxy-2 -piperidyl acetonyl -4 3H) quinazolinone The procedure of Example 12 is followed using 2.3 grams of 6,7-dibromo-4(3H)-quinaz0linone and 5 .0 grams of allyl-2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. The product is a dark brown syrup.

Example 45.-Preparation of 6,7-dibromo 3 [3-(3-hydroxy 2-piperidyl)acetonyl]4(3H)-quinazo1inone hydrobromide The product of Example 44 is hydrolyzed following the procedure of Example 13. Recrystallization from ethanol water (activated charcoal), gives 920 mg. of white crystals, melting point 250 dec.

Example 46 The nitrate salt of 6,8 dichloro-3-[3-(3-hydroxy-2- piperidy1)acetonyl]-4-(3H)-quinazolinone is obtained by taking up 50 mg. of the hydrobromide salt of this compound in 5 ml. of water and adding 3.0 grams of solid sodium nitrate with warming. After 48 hours at 25 a white microcrystalline product is obtained, melting point 235 dec.

Example 47.Preparation of 6-chloro-7-fluoro-4-(3H)- quinazolinone 6-fluoroisatin (16.5 g.) is stirred with 250 ml. of glacial acetic acid at and 13.5 g. of sulfuryl chloride added dropwise over 12 minutes. The deep red reaction mixture is heated under reflux for 2 hours and then cooled giving 12.6 g. of 5-chloro-6-fluoroisatin as an orange solid, melting point 239242. The isatin (10 g.) in ml. of N sodium hydroxide is oxidized with 20 ml. of 30% hydrogen peroxide giving 5.9 g. of 5-chloro-4-fluoroanthranilic acid. 4.75 g. of this anthranilic acid is cyclized as in Example 11 using 6 ml. of formamide and 3 ml. of dimethylformamide. Removal of the solvents under vacuum gives a gum which upon treatment with water becomes an off-white solid, melting point greater than 300.

Example 48.Preparation of 6-chloro-7-fluoro-3-[3-(1- allyloxycarbonyl 3 methoxy 2 piperidyl)acetonyl]- 4 3H) -quinazolinone The procedure of Example 12 is followed with 4.96 g. of 6-chloro-7-fiuoro-4(3H)-quinazolinone and 12.53 g. of allyl-2-(3-bromoacetonyl)-3-methoxy-1-piperidine carboxylate. The product is a dark brown syrup.

Example 49.Preparation of 6-chloro-7-fluoro-3-[3-(3- hydroxy 2 piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide The product of Example 48 is hydrolyzed according to the procedure of Example 13. Evaporation of the hydrobromic acid leaves a residue from which 1.75 g. of brown solid is obtained after several treatments with boiling absolute alcohol. Recrystallization from alcohol-water gives 650 mg. of solid, melting point 237 dec.

We claim:

1. A method of controlling coccidiosis in animals comprising orally administering to domestic animals an edible carrier containing an efiective amount of an anticoccidial of the formula:

wherein each of R R and R is individually selected from the group consisting of hydrogen, halogen, nitro, lower alkyl, phenyl and lower alkoxy, and R and R or R and R taken together, may form a benzo moiety, [wherein R is a member of the group consisting of hydrogen, halogen, nitro, benzo, lower alkyl, phenyl, and lower a1koxy,] R is a member of the group consisting of hydroxy, acetoxy, and lower alkoxy, and R is a member of the group consisting of hydrogen and lower alkenoxy-carbonyl, and the physiologically acceptable salts thereof.

2. A method according to claim 1 in which the quinazolinone is present to the extent of between about 0.5 and 10 parts per million and the edible carrier is an animal feed.

3. A method according to claim 1 in which the compound is 6-chloro-7-fiuoro-3-[3-(3-hydroxy-2-piperidyl)- acetonyl]-4(3H)-quinazolinone hydrobromide.

4. A method according to claim 1 in which the compound is 6,8-dichloro-3-[3-(3-hydroxy-2-piperidyl)-acetonyl]-4(3H)-quinazolinone hydrochloride.

5. A method according to claim 1 in which the compound is 6-phenyl-3-[3-(3-hydroxy-2-piperidyl)acetonyl]- 4(3H)-quinazolinone dihydrochloride.

6. A method according to claim 1 in which the compound is 6,7-dichloro-3-[3-(3-hydroxy-2-piperidyl)-acetonyl] -4( 3H) -quinazolinone hydrobromide.

7. A method according to claim 1 in which the compound is 6-chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl1- 4(3H)-quinazolinone.

8. A method according to claim 1 in which the compound is 6-chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl]- 4(3H)quinazolinone hydrobromide.

9. A method according to claim 1 in which the compound is 6-chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl1- 4(3H)-quinazolinone citrate.

10. A method according to claim 1 in which the compound is 6-nitro-3-[3-(3-hydroxy-2-piperidyl)acetonyll- 4(3H)-quinazolinone hydrochloride.

11. A method according to claim 1 in which the compound is 6,7,8 trichloro 3 [3 (3 hydroxy 2 piperidyl)-acetonyl]-4(3H)-quinazolinone hydrobromide.

12. A method according to claim 1 in which the compound is 7,8-dich1oro-3-[3-(3-hydroxy-2-piperidyl)-acetonyl]-4(3H)-quinazolinone hydrobromide.

13. A method according to claim 1 in which the compound is 8-chloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl]- 4(3H)-quinazolinone hydrobromide.

14. A method according to claim 1 in which the compound is 6,8-dichloro-3-[3-(3-hydroxy-2-piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide.

15. A method according to claim 1 in which the compound is 6,8-dibrom0-3-[3-(3-hydroxy-2-piperidyl)-acetonyl]-4(3H)-quinazolinone dihydrochloride.

16. A method according to claim 1 in which the compound is 6-methyl-7-chloro3-[3-(3-hydroxy-2-piperidyl)- acetonyl]-4( 3H -quinazolinone dihydrochloride.

17. A method according to claim 1 in which the compound is 6-methyl-7-chloro-3-[3-(3-hydroxy-2-piperidyl)- acetonyl]-4( 3H)-quinazolinone dihydrochloride.

18. A method according to claim 1 in which the compound is 7-br0mo-6-ch1oro-3-[3-(3-hydroxy-2-piperidyl)- acetonyl]-4(3H)-quinazolinone hydrobromide.

19. A method according to claim 1 in which the compound is 6-bromo-8-chloro-3-[3-(3-hydroxy-2-piperidyl)- acetonyl] -4(3H )-quinazolinone hydrobromide.

20. A method according to claim 1 in which the compound is 6,7-dibromo-3-[3-(3-hydroxy-2-piperidyl)acetonyl]-4(3H)-quinazolinone hydrobromide.

The following references, cited by the Examiner, are of record in the patented file of this patent or the original patent.

FOREIGN PATENTS 159,703 9/1951 Australia.

ALBERT T. MEYERS, Primary Examiner HOWARD M. ELLIS, Assistant Examiner US. Cl. X.R. 424-229 

